Attractene Transfection Reagent represents the next generation in lipid technology, ensuring highly efficient DNA transfection of eukaryotic cells. Attractene Reagent is a nonliposomal lipid that enables transfection of all adherent cells, including difficult-to-transfect cell types such as HaCaT, MonoMac6, and HCT116, and some suspension cell types (Jurkat, K562). It is also highly suitable for cotransfection of DNA with siRNA or miRNA mimics or inhibitors.

Attractene Transfection Reagent for up to 2640 transfections in 24-well plates

The Attractene Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Attractene Reagent outperforms alternative reagents.

DNA (pGFP which expresses green fluorescent protein) was transfected into the cell types indicated using Attractene Reagent according to the conditions recommended in the handbook. Transfection was also performed with [A] Reagent F using 2 reagent volumes recommended by the manufacturer or [B] Reagent L according to the manufacturer's recommendations, with and without the enhancer provided. Transfection efficiency was estimated by counting the number of fluorescent cells by FACS analysis. Transfection efficiency is expressed relative to efficiency using Attractene Reagent which was set at 100%.

Healthy cells after transfection using Attractene Reagent.

HepG2 cells were transfected with DNA (pGFP) using Attractene Transfection Reagent or Reagent L from another supplier according to the manufacturer's instructions. FACS analysis confirmed equal numbers of transfected cells in both cultures. Two days after transfection, cells were examined by light microscopy. [A] Cells transfected using Attractene Reagent were healthy and viable. [B] In contrast, cells transfected using Reagent L displayed high levels of cell death. [C] Cell viability was measured using a CellTiter-Blue assay (Promega). Viability was significantly lower in cells transfected using Reagent L compared to cells transfected using Attractene Reagent (set at 100%).

Flexible, rapid Fast-Forward Protocols.

Traditionally, cells are seeded the day before transfection. Using the Fast-Forward Protocol, seeding and transfection take place on the same day. This saves time and effort.

Effective knockdown after shRNA vector transfection.

HEK293 cells were [A] transfected with a plasmid expressing green fluorescent protein (pGFP) only, or [B] cotransfected with pGFP and a plasmid expressing an shRNA targeted against the green fluorescent protein gene (pGFP + pshGFP), or [C] cotransfected with pGFP and a negative control plasmid expressing a scrambled shRNA (pGFP + pNegative). After cotransfection of pGFP and the shRNA vector pshGFP, the green fluorescent protein was effectively silenced indicating effective cotransfection.

Attractene Reagent is a nonliposomal lipid which enables highly efficient DNA transfection of some suspension cell types (Jurkat, K562), and adherent cells, including difficult-to-transfect cell types such as HaCaT, MonoMac6, and HCT116. Attractene Reagent consistently provides higher transfection efficiency than alternative reagents under a range of experimental conditions. This feature allows flexible handling and ease of use for many experimental setups such as using automated platforms and transfection at high- or low-throughput levels. It is also highly suitable for cotransfection of DNA with siRNA or miRNA mimics or inhibitors. Using Attractene Reagent ensures exceptionally low cytotoxicity, which is critical to successful transfection experiments.

Procedure

Attractene Transfection Reagent is highly suited for rapid fast-forward DNA transfection. In Fast-Forward Protocols, cells are seeded and transfected on the same day (see figure "Flexible, rapid Fast-Forward Protocols"). This is quicker, saves labor, and increases experimental flexibility compared to protocols where cells are seeded the day before transfection.

Cell type-specific protocols at the TransFect Protocol Database

In addition to the protocols provided in the Attractene Transfection Reagent Handbook, you can find protocols to suit your cell type and plate/dish format using QIAGEN's TransFect Protocol Database. The database provides exactly the protocol needed, saving time and effort. Simply enter the cell type and nucleic acid of interest to receive a QIAGEN transfection protocol to print out or download in convenient PDF format. Use of the TransFect Protocol Database is free of charge and no registration is required.

Applications

Attractene Transfection Reagent can be used for a range of transfection applications, including transient or stable transfection, or cotransfection of multiple DNA constructs, and cotransfection of DNA with siRNA or miRNA mimics or inhibitors. Attractene Transfection Reagent can be used in: